Molecular genetics of the Drosophila eyes absent gene

The Drosophila compound eye has provided a genetic approach to understanding the specification of cell fates during differentiation. The eye is made up of some 750 repeated units or ommatidia, arranged in a lattice. The cellular composition of each ommatidium is identical. The arrangement of the lattice and the specification of cell fates in each ommatidium are thought to occur in development through cellular interactions with the local environment. Many mutations have been studied that disrupt the proper patterning and cell fating in the eye. The eyes absent (eya) mutation, the subject of this thesis, was chosen because of its eyeless phenotype. In eya mutants, eye progenitor cells undergo programmed cell death before the onset of patterning has occurred. The molecular genetic analysis of the gene is presented.

The eye arises from the larval eye-antennal imaginal disc. During the third larval instar, a wave of differentiation progresses across the disc, marked by a furrow. Anterior to the furrow, proliferating cells are found in apparent disarray. Posterior to the furrow, clusters of differentiating cells can be discerned, that correspond to the ommatidia of the adult eye. Analysis of an allelic series of eya mutants in comparison to wild type revealed the presence of a selection point: a wave of programmed cell death that normally precedes the furrow. In eya mutants, an excessive number of eye progenitor cells die at this selection point, suggesting the eya gene influences the distribution of cells between fates of death and differentiation.

In addition to its role in the eye, the eya gene has an embryonic function. The eye function is autonomous to the eye progenitor cells. Molecular maps of the eye and embryonic phenotypes are different. Therefore, the function of eya in the eye can be treated independently of the embryonic function. Cloning of the gene reveals two cDNA's that are identical except for the use of an alternatively-spliced 5' exon. The predicted protein products differ only at the N-termini. Sequence analysis shows these two proteins to be the first of their kind to be isolated. Trangenic studies using the two cDNA's show that either gene product is able to rescue the eye phenotype of eya mutants.

The eya gene exhibits interallelic complementation. This interaction is an example of an "allelic position effect": an interaction that depends on the relative position in the genome of the two alleles, which is thought to be mediated by chromosomal pairing. The interaction at eya is essentially identical to a phenomenon known as transvection, which is an allelic position effect that is sensitive to certain kinds of chromosomal rearrangements. A current model for the mechanism of transvection is the trans action of gene regulatory regions. The eya locus is particularly well suited for the study of transvection because the mutant phenotypes can be quantified by scoring the size of the eye.

The molecular genetic analysis of eya provides a system for uncovering mechanisms underlying differentiation, developmentally regulated programmed cell death, and gene regulation.

New technologies driving decade-bandwidth radio astronomy : quad-ridged flared horn and compound-semiconductor LNAs

Among the branches of astronomy, radio astronomy is unique in that it spans the largest portion of the electromagnetic spectrum, e.g., from about 10 MHz to 300 GHz. On the other hand, due to scientific priorities as well as technological limitations, radio astronomy receivers have traditionally covered only about an octave bandwidth. This approach of "one specialized receiver for one primary science goal" is, however, not only becoming too expensive for next-generation radio telescopes comprising thousands of small antennas, but also is inadequate to answer some of the scientific questions of today which require simultaneous coverage of very large bandwidths.

This thesis presents significant improvements on the state of the art of two key receiver components in pursuit of decade-bandwidth radio astronomy: 1) reflector feed antennas; 2) low-noise amplifiers on compound-semiconductor technologies. The first part of this thesis introduces the quadruple-ridged flared horn, a flexible, dual linear-polarization reflector feed antenna that achieves 5:1-7:1 frequency bandwidths while maintaining near-constant beamwidth. The horn is unique in that it is the only wideband feed antenna suitable for radio astronomy that: 1) can be designed to have nominal 10 dB beamwidth between 30 and 150 degrees; 2) requires one single-ended 50 Ohm low-noise amplifier per polarization. Design, analysis, and measurements of several quad-ridged horns are presented to demonstrate its feasibility and flexibility.

The second part of the thesis focuses on modeling and measurements of discrete high-electron mobility transistors (HEMTs) and their applications in wideband, extremely low-noise amplifiers. The transistors and microwave monolithic integrated circuit low-noise amplifiers described herein have been fabricated on two state-of-the-art HEMT processes: 1) 35 nm indium phosphide; 2) 70 nm gallium arsenide. DC and microwave performance of transistors from both processes at room and cryogenic temperatures are included, as well as first-reported measurements of detailed noise characterization of the sub-micron HEMTs at both temperatures. Design and measurements of two low-noise amplifiers covering 1--20 and 8—50 GHz fabricated on both processes are also provided, which show that the 1--20 GHz amplifier improves the state of the art in cryogenic noise and bandwidth, while the 8--50 GHz amplifier achieves noise performance only slightly worse than the best published results but does so with nearly a decade bandwidth.

Molecular neurogenetics of eye development in Drosophila

The compound eye of Drosophila melanogaster begins to differentiate during the late third larval instar in the eye-antennal imaginal disc. A wave of morphogenesis crosses the disc from posterior to anterior, leaving behind precisely patterned clusters of photoreceptor cells and accessory cells that will constitute the adult ommatidia of the retina. By the analysis of genetically mosaic eyes, it appears that any cell in the eye disc can adopt the characteristics of any one of the different cell types found in the mature eye, including photoreceptor cells and non-neuronal accessory cells such as cone cells. Therefore, cells within the prospective retinal epithelium assume different fates presumably via information present in the environment. The sevenless^+ (sev^+) gene appears to play a role in the expression of one of the possible fates, since the mutant phenotype is the lack of one of the pattern elements, namely, photoreceptor cell R7. The sev^+ gene product had been shown to be required during development of the eye, and had also been shown in genetic mosaics to be autonomous to presumptive R7. As a means of better understanding the pathway instructing the differentiation R7, the gene and its protein product were characterized.

The sev+ gene was cloned by P-element transposon tagging, and was found to encode an 8.2 kb transcript expressed in developing eye discs and adult heads. By raising monoclonal antibodies (MAbs) against a sev^+- β-galactosidase fusion protein, the expression of the protein in the eye disc was localized by immuno-electronmicroscopy. The protein localizes to the apical cell membranes and microvilli of cells in the eye disc epithelium. It appears during development at a time coincident with the initial formation of clusters, and in all the developing photoreceptors and accessory cone cells at a time prior to the overt differentiation of R7. This result is consistent with the pluripotency of cells in the eye disc. Its localization in the membranes suggests that it may receive information directing the development of R7. Its localization in the apical membranes and microvilli is away from the bulk of the cell contacts, which have been cited as a likely regions for information presentation and processing. Biochemical characterization of the sev^+ protein will be necessary to describe further its role in development.

Other mutations in Drosophila have eye phenotypes. These were analyzed to find which ones affected the initial patterning of cells in the eye disc, in order to identify other genes, like sev, whose gene products may be involved in generating the pattern. The adult eye phenotypes ranged from severe reduction of the eye, to variable numbers of photoreceptor cells per ommatidium, to sub de defects in the organization of the supporting cells. Developing eye discs from the different strains were screened using a panel of MAbs, which highlight various developmental stages. Two identified matrix elements in and anterior to the furrow, while others identified the developing ommatidia themselves, like the anti-sev MAb. Mutation phenotypes were shown to appear at many stages of development. Some mutations seem to affect the precursor cells, others, the setting up of the pattern, and still others, the maintenance of the pattern. Thus, additional genes have now been identified that may function to support the development of a complex pattern.

Schottky barriers on compound semiconductors: I. HgSe highly electronegative contacts. II. Au Schottky barriers on n-Ga_(1-x)Al_xAs

I. HgSe is deposited on various semiconductors, forming a semimetal/semiconductor "Schottky barrier" structure. Polycrystalline, evaporated HgSe produces larger Schottky barrier heights on n-type semiconductors than does Au, the most electronegative of the elemental metals. The barrier heights are about 0.5 eV greater than those of Au on ionic semiconductors such as ZnS, and 0.1 to 0.2 eV greater for more covalently bonded semiconductors. A novel structure,which is both a lattice matched heterostructure and a Schottky barrier, is fabricated by epitaxial growth of HgSe on CdSe using hydrogen transport CVD. The Schottky barrier height for this structure is 0.73 ± 0.02 eV, as measured by the photoresponse method. This uncertainty is unusually small; and the magnitude is greater by about a quarter volt than is achievable with Au, in qualitative agreement with ionization potential arguments.

II . The Schottky barrier height of Au on chemically etched n-Ga_1-x Al_xAs was measured as a function of x. As x increases, the barrier height rises to a value of about 1.2 eV at x ≈ 0.45 , then decreases to about 1.0 eV as x approaches 0.83. The barrier height deviates in a linear way from the value predicted by the "common anion" rule as the AlAs mole fraction increases. This behavior is related to chemical reactivity of the Ga_1-x Al_xAs surface.